Phylogenetically Distant Viruses Use the Same BH3-Only Protein Puma to Trigger Bax/Bak-Dependent Apoptosis of Infected Mouse and Human Cells

Emanuela Papaianni; Souhayla El Maadidi; Andrea Schejtman; Simon Neumann; Ulrich Maurer; Francesca Marino-Merlo; Antonio Mastino; Christoph Borner

doi:10.1371/journal.pone.0126645

Phylogenetically Distant Viruses Use the Same BH3-Only Protein Puma to Trigger Bax/Bak-Dependent Apoptosis of Infected Mouse and Human Cells

PLoS One. 2015 Jun 1;10(6):e0126645. doi: 10.1371/journal.pone.0126645. eCollection 2015.

Authors

Emanuela Papaianni¹, Souhayla El Maadidi², Andrea Schejtman³, Simon Neumann⁴, Ulrich Maurer⁵, Francesca Marino-Merlo⁶, Antonio Mastino⁷, Christoph Borner⁵

Affiliations

¹ Department of Biological and Environmental Sciences, University of Messina, Via F. Stagno d'Alcontres 31, I-98166, Messina, Italy; The Institute of Translational Pharmacology, CNR, Via Fosso del Cavaliere 100, I-00133, Rome, Italy; Institute of Molecular Medicine and Cell Research, Albert Ludwigs University of Freiburg, Stefan Meier Strasse 17, D-79104, Freiburg, Germany.
² Institute of Molecular Medicine and Cell Research, Albert Ludwigs University of Freiburg, Stefan Meier Strasse 17, D-79104, Freiburg, Germany; Faculty of Biology, Albert Ludwigs University of Freiburg, Schänzlestrasse 1, D-79104, Freiburg, Germany.
³ Institute of Molecular Medicine and Cell Research, Albert Ludwigs University of Freiburg, Stefan Meier Strasse 17, D-79104, Freiburg, Germany; IMBS Program between Albert Ludwigs University of Freiburg, Freiburg, Germany, and University of Buenos Aires, Buenos Aires, Argentina.
⁴ Institute of Molecular Medicine and Cell Research, Albert Ludwigs University of Freiburg, Stefan Meier Strasse 17, D-79104, Freiburg, Germany.
⁵ Institute of Molecular Medicine and Cell Research, Albert Ludwigs University of Freiburg, Stefan Meier Strasse 17, D-79104, Freiburg, Germany; Spemann Graduate School of Biology and Medicine (SGBM), Albert Ludwigs University of Freiburg, Albertstrasse 19a, D-79104, Freiburg, Germany; BIOSS, Centre for Biological Signaling Studies, Hebelstrasse 2, D-79104, Freiburg, Germany.
⁶ Department of Biological and Environmental Sciences, University of Messina, Via F. Stagno d'Alcontres 31, I-98166, Messina, Italy.
⁷ Department of Biological and Environmental Sciences, University of Messina, Via F. Stagno d'Alcontres 31, I-98166, Messina, Italy; The Institute of Translational Pharmacology, CNR, Via Fosso del Cavaliere 100, I-00133, Rome, Italy.

Abstract

Viruses can trigger apoptosis of infected host cells if not counteracted by cellular or viral anti-apoptotic proteins. These protective proteins either inhibit the activation of caspases or they act as Bcl-2 homologs to prevent Bax/Bak-mediated outer mitochondrial membrane permeabilization (MOMP). The exact mechanism by which viruses trigger MOMP has however remained enigmatic. Here we use two distinct types of viruses, a double stranded DNA virus, herpes simplex virus-1 (HSV-1) and a positive sense, single stranded RNA virus, Semliki Forest virus (SFV) to show that the BH3-only protein Puma is the major mediator of virus-induced Bax/Bak activation and MOMP induction. Indeed, when Puma was genetically deleted or downregulated by shRNA, mouse embryonic fibroblasts and IL-3-dependent monocytes as well as human colon carcinoma cells were as resistant to virus-induced apoptosis as their Bax/Bak double deficient counterparts (Bax/Bak-/-). Puma protein expression started to augment after 2 h postinfection with both viruses. Puma mRNA levels increased as well, but this occurred after apoptosis initiation (MOMP) because it was blocked in cells lacking Bax/Bak or overexpressing Bcl-xL. Moreover, none of the classical Puma transcription factors such as p53, p73 or p65 NFκB were involved in HSV-1-induced apoptosis. Our data suggest that viruses use a Puma protein-dependent mechanism to trigger MOMP and apoptosis in host cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis Regulatory Proteins / genetics
Apoptosis Regulatory Proteins / metabolism*
Apoptosis*
Caspase 3 / metabolism
Cytochromes c / metabolism
Enzyme Activation
Fas Ligand Protein / metabolism
Fibroblasts / metabolism
HCT116 Cells
Herpesvirus 1, Human / metabolism*
Humans
Mice
Nuclear Pore Complex Proteins
Phylogeny*
Proto-Oncogene Proteins / genetics
Proto-Oncogene Proteins / metabolism*
RNA, Messenger / genetics
RNA, Messenger / metabolism
RNA-Binding Proteins
Receptor-Interacting Protein Serine-Threonine Kinases / metabolism
Semliki forest virus / metabolism*
TNF-Related Apoptosis-Inducing Ligand / metabolism
Tumor Suppressor Proteins / genetics
Tumor Suppressor Proteins / metabolism*
U937 Cells
Virus Replication
bcl-2 Homologous Antagonist-Killer Protein / metabolism*
bcl-2-Associated X Protein / metabolism*

Substances

AGFG1 protein, human
Apoptosis Regulatory Proteins
BBC3 protein, human
Fas Ligand Protein
Nuclear Pore Complex Proteins
PUMA protein, mouse
Proto-Oncogene Proteins
RNA, Messenger
RNA-Binding Proteins
TNF-Related Apoptosis-Inducing Ligand
Tumor Suppressor Proteins
bcl-2 Homologous Antagonist-Killer Protein
bcl-2-Associated X Protein
Cytochromes c
RIPK3 protein, human
Receptor-Interacting Protein Serine-Threonine Kinases
Caspase 3

Grants and funding

This work was supported by the Spemann Graduate School of Biology and Medicine (SGBM, GSC-4) and the Centre for Biological Signaling Studies (BIOSS, EXC-294) (to CB and UM), both funded by the Excellence Initiative of the German Federal and State Governments, Germany as well as by the Research Group FOR2036 funded by the German Research Foundation (DFG) (to CB and SN). AM, FMM and EP were financed by the Italian Ministry of University and Research, Projects of National Interest. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.